MXPA06001145A - Thiomorpholino steroid compounds, the use thereof for the preparation of meiosis-regulating medicaments and method for the preparation thereof - Google Patents

Abstract

The present invention relates to thiomorpholino steroid compounds of general formula (I), which may advantageously be employed to stimulate meiosis in human oocytes, the steroid being specifically characterized by a thiomorpholino moiety bonded to C17 of the steroid skeleton via an alkylen spacer.

Description

COMPOUNDS TIOMORFOLINO ESTEROIDES, ITS USE FOR THE PREPARATION OF REGULATORY MEDICINES OF THE MEIOSIS AND METHOD FOR PREPARING THEM PREVIOUS ART Meiosis is the unique and final event of the germ cells, and sexual reproduction is based on it. Meiosis comprises two meiotic divisions. During the first division, the exchange between the maternal and paternal genes takes place, before the pairs of chromosomes between the daughter cells are separated. These contain only half the amount (1 n) of chromosomes and 2c of DNA. The second meiotic division proceeds without DNA synthesis. Accordingly, this division results in the formation of haploid germ cells with only 1c of DNA. The meiotic process is similar in male and female germ cells, but the timing and differentiation process, which leads to eggs and sperm, differ profoundly. All female germ cells enter the prophase of the first meiotic division at an early stage of their life, commonly before birth, but they stop as occidentals in a late stage of the prophase (state of death) until ovulation occurs after puberty. Therefore, from a moment early in life, the female has a reserve of oocytes, which is used until it is exhausted. Meiosis in females is not complete until after fertilization, and results only in one egg and two abortive polar bodies per germ cell. In contrast, only some of the male germ cells enter meiosis at puberty and leave the germ cell population over the course of life. Once initiated, meiosis in the male cell proceeds without significant delays, and produces four sperm cells. Little is known about the mechanisms that control the onset of meiosis in the male and female. The newer studies indicate that follicular purines, hypoxanthine and adenosine, could be responsible for the arrest of meiosis in the oocyte [S.M. Downs et al., Dev. Biol., 82, 454-458 (1985); J.J. Epplg. et al., Dev. Biol., 119, 313-321 (1986); YE. Downs, Mol. Reprod. Dev., 35, 82-94 (1993)]. The presence of a diffusible substance that regulates meiosis was first described by Byskov et al. in a mouse fetal gonad culture system [A.G. Byskov et al., Dev. Biol., 52, 193-200 (1976)]. There is a meiosis-activating substance (MAS) that is secreted by the fetal ovary of the mouse in which meiosis occurs, and a substance that prevents meiosis (MPS), which is released from the testes in morphological differentiation with germ cells. meiotic at rest. It was suggested that the relative concentrations of MAS and MPS regulated the initiation, arrest and continuation of meiosis in male and female germ cells [A.G. Byskov et al. in: The Physiology of Reproduction (edited by E. Knobil and J.D. Neill), Raven Press, New York (1994)].

Clearly, if it is possible to regulate meiosis, it is possible to control reproduction. In a recent article [A.G. Byskov et al., Nature, 374, 559-562 (1995)] describes the isolation of certain stereos. These steles were isolated from bull testes and follicular fluid, and activated meiosis of the oocyte [T-MAS (sterol activator of meiosis in the testes) and FF-MAS (sterol activator of meiosis in the follicular fluid): 4,4-dimethyl-5a-cholesta-8,14,24-trien-3ß-ol]. It was also shown that micromolar concentrations of synthetic FF-MAS were able to induce the continuation of meiosis in a dose-dependent manner, in rat oocytes arrested by the phosphodiesterase inhibitor IBMX (3-isobutyl-1-methyl xanthine). ) [C. Hegele-Hartung et al., Biol. Reprod., 64, 418-424 (2001)]. It was shown that it was possible to observe this effect when CEO (oocytes encased in clusters) and DO (naked oocytes) were cultured in vitro in the presence of FF-MAS. Other substances that regulate meiosis are described in the following documents. WO 98/52965 A1 discloses 20-aralkyl-5α-pregnane derivatives that activate meiosis. WO 00/68245 A1 discloses spheroidal compounds capable of inhibiting meiosis, so that these compounds are useful as contraceptives in females and males. These compounds are primarily unsaturated cholestane derivatives, characterized by a 3β-hydrogen atom attached to the C 14 carbon atom of the cholestane backbone. In WO 96/00235 A1, meiosis-inducing esterals known as intermediates in cholesterol biosynthesis are described; as well as certain synthetic material related from the structural point of view. It has been discovered that these substances regulate meiosis. Similar to cholesterol, these esterals are provided with a C17 side chain in the sterol backbone, and also contain at least one double bond D7, D8 or D8 (14). In WO 96/27658 A1 a method for stimulating meiosis of a germ cell is described, which comprises administering to the cell in vivo, ex vivo or in vitro an effective amount of a compound, which causes the accumulation of an activating substance of the endogenous meiosis to a level at which meiosis is induced. These compounds that cause the accumulation of the activating substance of meiosis include amphotericin B, aminoguanidine, Sß.Sa.dβ-trihydroxycholestane, melatonin, 6-chloromelatonin and 5-methoxytryptamine, as well as other derivatives and agonists thereof. It is also reported that the meiosis activating substances include, among others, 5a-cholestan-3ß-ol, D-homo-cholesta-8,14-dien-3ß-ol and 22,25-diazacolestrol, 25-aza-24 , 25-dihydrolanosterol, 24,25-iminolano-sterol, 23 and 24-azacolestrol, and also 25-azacolestanol derivatives. WO 97/00884 A1 and WO 98/28323 A1 describe substances that can be used to stimulate meiosis in vitro, in vivo or ex vivo. Accordingly, the described compounds are agonists of meiosis activating substances of natural occurrence, so they can be used in the treatment of infertility due to insufficient stimulation of meiosis in females and males. This document also describes some compounds that can be antagonists of naturally occurring meiosis activating substances, so that these compounds may be suitable for use as contraceptives. The compounds described include, among others, 5α-cholest-8-ene-3β-oles and 5α-cholest-8,14-dien-3β-ols, which, among others, may comprise an amino group in the side chain, to C17 of the cholesterol skeleton, where the amino group is attached to the sterol backbone via a C4 spacer. There is a C 1 -C 4 alkyl or a C 3 -C 6 -cycloalkyl bonded to the amino group. In addition, WO 99/58549 A1 describes sterol derivatives that are effective in regulating meiosis. It is established that these have the ability to alleviate infertility in females and males, particularly in humans. Sterol derivatives that are effective as regulatory substances include, but are not limited to, (20R) -20-methyl-23-dimethylamino-5a-pregna-8,14-diene-3ß-ol, (20R) -20-methyl-23 -dimethylamino-5a-pregna-5,7-dien-3ß-ol, 4,4-dimethyl-24-phenylamino-5a-cola-8,14-dien-3ß-ol, 4,4-dimetyl-24 - (N, N-dimethylamino) -24-cyano-5a-cholesta-8,14-diene-3ß-ol, and in addition, a variety of 24-oic acid amides of stellate having one or more double bonds in the Sterol skeleton.

WO 02/079220 A2 discloses steroid compounds that are capable of regulating meiosis, wherein these compounds are primarily stereos having a portion of aminomethyl or aminoethyl linked to C20. The amino group can be, for example, a heterocyclic ring containing nitrogen, more specifically a piperidine ring. An example of these compounds is (20S) -20 - [(piperidin-1-yl) methyl] -4,4-dimethyl-5a-pregna-8, 14-dien-3ß-ol. In general, these compounds have an effect of stimulation of meiosis in oocytes, especially in CEO (= oocytes enclosed in clusters). In a specific example, a thiomorpholino compound is disclosed. The unsaturated sterol derivatives having an amino group at C17 of the side chain have also been described by J.J. Sheets and L.E. Vickery in: "Active Site-directed Inhibitors of Cytochrome P-450scc" in J. Biol. Chem., Vol.258 (19), 1983, pages 11446-11452, with reference to the effect of these esterases on cytochrome P-450 , which cuts the side chain of bovine adenocortical cholesterol (P-450 scc). This document describes, among others, 22-amino-23,24-bis-nor-col-5-en-3ß-ol and 23-amino-24-norcol-5-en-3ß-ol. Other unsaturated derivatives having an amino group on the C17 side chain have been described in A.T. Mangla and W.D. Nes in: "Sterol C-methyl Transferase from Prototheca wickerhamii, Mechanism, Sterol Specificity and Inhibition", Bioorg. And Med. Chem. (2000), 8 (5), 925-936. This document describes, among others, 23-aza-zymosterol.

It has been discovered that, when the meiosis regulatory components described above are used, it is possible to cause the continuation of meiosis in naked oocytes in vitro. However, many of these compounds had only marginal effectiveness when meiosis was stimulated in oocytes surrounded by granulosa cells (CEO). In addition, the rates of in vitro fertilization and re-transfer implantation, and also the number of live fetuses at the end of pregnancy, are not sufficiently high. The description of the above documents is incorporated by reference. SUMMARY OF THE INVENTION The invention relates to steroid-containing thiomorpholino compounds with pharmaceutical activity, with pharmaceutical compositions comprising these compounds, with the use of these compounds for the preparation of a pharmaceutical composition suitable for regulating the reproduction, especially meiosis, of a contraceptive, or as pro-fertility drug, with a method for regulating reproduction, for example, meiosis, with a method to improve the possibility of an oocyte developing in a mammal using these compounds, as well as with a method for the preparation of ( 20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-5a-pregna-8,14-dien-3ß-ol.

DETAILED DESCRIPTION OF THE INVENTION An object of the present invention is to discover substances that are useful for regulating reproduction, in particular meiosis, in females and males, especially in mammals, and more specifically in humans. Another object of the present invention is to provide a new pharmaceutical composition comprising the new substances. Another preferred object of the present invention is to provide the use of the new substances for the preparation of an appropriate pharmaceutical composition for regulating reproduction, especially meiosis. Another preferred object of the present invention is to provide a new method for regulating reproduction, for example, meiosis. Another object of the present invention is to provide a method for treating human infertility. Another object of the present invention is to improve the maturation of human oocytes. Still another object of the present invention is to improve the synchronization of the nuclear, cytoplasmic and / or membranous maturation of the oocyte. Still another object of the present invention is to improve the fertility of oocytes.

Still another object of the present invention is to improve the implantation rate of oocytes by maturation and in vitro fertilization in humans. Still another object of the present invention is to reduce the incidence of pre-embryos with chromosomal abnormalities (aneuploidy). Still another object of the present invention is to improve the cutting rate of human pre-embryos. Still another object of the present invention is to improve the quality of human pre-embryos. Another object of the present invention is to provide a method for the preparation of the new substances. According to the present invention, the steroid thiomorpholino compounds of the general formula I can be advantageously used in the regulation of reproduction, for example, meiosis, in mammals, for example, in females and males, and in particular in humans: where, in the I 'portion of the compound I r each link between C5 and C6, between C6 and C7, between C7 and C8, between C8 and C9, between C8 and C14 and between C14 and C15, independently, it is a simple link or a double link, where at least one of these links is a double bond, and each carbon atom C5, C6, C7, C8, C9, C14 and C15 is attached to each atom of C adjacent to at most one double bond, bearing in mind that there should not be a double bond in the skeleton of the steroid exclusively between C5 and C6 (where this latter condition indicates that compounds having exclusively a double bond D5 are not included in the present invention), and where R4 and R4, independently, are selected from the group comprising hydrogen and methyl. The portion with the general formula I 'in the steroid compound according to the present invention preferably comprises a double bond between C8 and C14, or two conjugated double bonds, preferably two double bonds between C8 and C9, and between C14 and C15, or two double bonds between C5 and C6, and between C7 and C8. R4 and R4 are preferably the same radicals, that is, they are both hydrogen or both methyl.

The rest of the hydrogen atoms may be attached to C1, C2, C5, C6, C7, optionally C8, C9, C11, C12, C14, C15, C16 and C17, depending on whether the respective C atoms are part of a double link or not. In C10, C13 and C18 there are methyl groups attached to the skeleton of the steroid and the side chain, respectively. More preferably, according to the present invention, the steroid compound is selected from the group comprising: (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-d, methyl-5 -pregna-8 , 14-dien-3ß-ol: Compound IA (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-5a-pregna-8 (14) -en-3ß-ol: Compound IB (20S) -20 - [(thiomorpholin-4-yl) methyl] -5a-pregna-8,14-dien-3ß-ol: Compound IC (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-pregna-5,7-diene-3ß-ol: Compound ID (20S) -20 - [(thiomorpholin-4-yl) methyl] -5a-pregna-8 (14) -en-3ß-ol: Compound IE The most preferred thiomorpholino compound according to the present invention is (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-d-methyl-5a-pregna-8,14-dien -3ß-ol, which has the chemical formula IA. Accordingly, optimal performance is obtained with respect to the induction of maturation in a follicular culture system with thiomorpholino derivatives if the steroid backbone comprises a double bond system D8,14, and if R 4 and R 4 'are methyl (compound IA).

The new steroid compounds have a number of chiral centers, so that these compounds exist in various isomeric forms. All of these isomeric forms are within the scope of the present invention, unless otherwise indicated in this documentation. Surprisingly, it has been discovered that the compounds according to the present invention have a strong effect of stimulating meiosis in oocytes, especially in CEO, although these compounds present great structural differences with respect to the sterol FF-MAS. In this regard, the compounds of this invention are superior to this meiosis regulatory substance previously described [for example: A.G. Byskov et al., Nature, 374, 559-562 (1995)]. Preferred compounds of general formula I are those that induce the degradation of the germinal vesicle by at least 40%, preferably at least 60% and especially at least 80%, when evaluated in an oocyte test, as described further forward in this documentation. The compounds according to the present invention are superior to the compounds previously described in a second aspect: while FF-MAS is not capable of inducing maturation in a follicle culture system, the compounds of the present invention are capable of to activate meiosis in this situation. Compared to the compounds described in WO 02/079220 A2, the thio-morpholino steroid compounds of the present invention exhibit an even stronger stimulation activity, especially in in vitro fertilization, and a higher rate of repletion. successful transfer to the tubes of a female after in vitro fertilization. In addition, the new compounds have superior properties regarding the amount of live fetuses at the end of pregnancy, if the re-transfer of stimulated oocytes has been effected. Moreover, the solubility of the compounds according to the present invention in water is better than the solubility of the compounds of the prior art. It has also been found that the growth of the blastocysts is stimulated in a superior way, which can be demonstrated in mice. The above advantages of the novel compounds also apply to the thio-morpholin compounds according to the present invention, when compared to the more efficient compounds described in WO 02/079200 A2, especially at (20S) -20- [ (piperidin-1-yl) methyl] -4,4-dimethyl-5a-pregna-8,14-diene-3ß-ol (compound No. 2 in this document). For this reason, the new steroid compounds can be used, for example, for in vivo use, as well as for a use other than in vivo, which especially comprises in vitro use. Steroid compounds are especially suitable for the in vitro and in vivo fertilization of mammals, especially humans. The outstanding properties of the new compounds can be attributed to the combination of structural characteristics of the compounds, ie, primarily the position of the double bonds in the steroid backbone and the thiomorpholino group in the side chain attached to the carbon atom C17 in the skeleton of the steroid through a methylene spacer and the C20-R20 group. Preferably, the compounds acceptable for the pharmaceutical use of the present invention are salts of the steroid compounds of general formula I. Examples of these salts are detailed in Journal of Pharmaceutical Science, 66, 2 et seq. (1977), which is incorporated in this documentation as a reference. Examples of these salts include salts of organic acids, such as formic acid, fumaric acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, succinic acid, malic acid, tartaric acid, citric acid, benzoic acid, salicylic acid, methanesulfonic acid and the like. Suitable inorganic acids to form the salts acceptable for pharmaceutical use include hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid and the like. Another object of the present invention is pharmaceutical compositions comprising at least one steroid thiomorpholino compound of general formula I and at least one excipient acceptable for pharmaceutical use, selected from the group of excipients well known in the art. For example, the excipient may be at least one carrier, a diluent, an absorption enhancer, a preservative, a buffer, an agent for adjusting the osmotic pressure and the rheology of the medicament if the pharmaceutical composition is a liquid, at least one surfactant, a solvent, a tablet disintegrating agent, microcapsules, a filler, a slip additive, a colorant, a flavoring and other ingredients. These substances are commonly used in the art. The steroid thiomorpholino compounds according to the present invention are preferably included in the pharmaceutical compositions in an effective amount. Examples of solid carriers include magnesium carbonate, magnesium stearate, dextrin, lactose, sugar, talc, gelatin, pectin, starch, silica gel, tragacanth, methylcellulose, sodium carboxymethyl cellulose, low melting waxes, and cocoa butter. . Liquid compositions include solutions, suspensions and emulsions, which can be administered, for example, orally, by nasal administration or as an ointment. These liquid compositions may also be suitable for injection or for use in combination with an ex vivo or in vivo application. For oral administration, the liquid may contain an oil and / or a lipophilic substance acceptable for pharmaceutical use, a surfactant and / or a solvent miscible with water. In this aspect, WO 97/21440 A1 can be consulted. The liquid compositions may also contain other ingredients commonly used in the art, some of which have been mentioned in the above list. In addition, a transdermal administration composition of a compound of the present invention may be provided in the form of a patch.

A composition for nasal administration may be provided in the form of a nasal spray, in liquid form or in powder form. In order to improve the bioavailability of the steroid thiomorpholino compound, these compounds can also be formulated as cyclodextrin clathrates. For this purpose, the compounds are combined with a, β or β-cyclodextrin, or derivatives thereof. Ointments, ointments, lotions and other externally administered liquids must be in a condition such that the steroid thiomorpholino compounds of the present invention can be administered in a sufficient amount to a subject in whom it is necessary to regulate meiosis. For this purpose, the drug contains the above excipients to regulate the rheology of the drug, and other additives, other substances to improve the ability to permeate the skin and protective substances, such as conditioners and moisture regulators. The medicament may also contain other active agents to improve or regulate the effectiveness of the thiomorpholino spheroidal compounds, or to produce other desired effects with the medicament. For parenteral administration, the steroid compounds can be dissolved or suspended in a diluent acceptable for pharmaceutical use. Oils are commonly used in combination with solvents, surfactants, suspending or emulsifying agents, for example, olive oil, peanut oil, soybean oil, castor oil. Any liquid vehicle can be used for the preparation of an injectable drug. These liquids commonly also contain agents for regulating viscosity, as well as agents for regulating the isotonicity of the liquid. In addition, the steroid thiomorpholino compound can be administered as an injectable reservoir or as an implant, which for example can be administered subcutaneously, so that the delayed release of the steroid thiomorpholino compounds is made possible. For this purpose, various techniques may be employed, for example, administration through a reservoir, including a membrane containing the active compound, or administration through a slow dissolution reservoir. For example, the implanted ones may contain biodegradable polymers in the biological field or synthetic silicones as an inert material. The used dose of the steroid thiomorpholino compound will be determined by a physician, and will depend, among others, on the particular steroid compound employed, the route of administration and the purpose of use. In general, the pharmaceutical compositions of the present invention are prepared by intimately associating the active compound with the liquid or solid auxiliary ingredients, and then, if necessary, giving the product the form of the desired formulation.

Typically, no more than 3000 mg, preferably no more than 350 mg, and in some preferred instances no more than 30 mg of the steroid compounds will be administered to mammals, eg, humans, per day. The present invention also relates to the use of the steroid thiomorpholino compounds of general formula I for the preparation of a composition useful for regulating reproduction, for example, meiosis. This composition is preferably applied as a medicine. In addition, the present invention relates to the use of the new steroid thiomorpholino compounds of general formula I in the preparation of a contraceptive or a pro-fertility drug. In addition, the present invention relates to the use of the steroid thiomorpholino compound of general formula I for an application that is not in vivo. The present invention also relates to a method for regulating reproduction, for example, meiosis, which comprises administering to a subject in need of such regulation an effective amount of at least one steroid thiomorpholino compound of general formula I. Further, the present invention is related to a method for improving the possibility of an oocyte developing in a mammal, comprising contacting an oocyte extracted from the mammal with the steroid thiomorpholino compound of general formula I.

In the present documentation, the regulation of reproduction is used, for example, to indicate that the compounds according to the present invention are especially suitable for stimulating reproduction, for example, meiosis, in a mammal, especially in humans, of oocytes, so that these compounds, which are agonistic analogs of a naturally occurring meiosis activating substance (FF-MAS), can be used in the treatment of infertility due to insufficient stimulation of meiosis in females and males.

The route of administration of the compositions containing a compound of the present invention can be any route that allows the active steroid compound to be transported effectively to its site of action. Accordingly, when the steroid compounds are administered to a mammal, they are conveniently provided in the form of a pharmaceutical composition comprising at least one steroid thiomorpholino compound according to the present invention, in combination with a vehicle acceptable for pharmaceutical use. For oral use, these compositions preferably take the form of tablets or capsules. The steroid thiomorpholino compounds can be synthesized analogously to the preparation of known compounds. Accordingly, the synthesis of the steroid compounds of formula I can be carried out in accordance with the well established pathways described in the extensive literature on steroids and esterases. The following literature can be used as a key source for the synthesis: L.F. Fieser & M. Fieser: Steroids, Reinhold Publishing Corporation, NY, 1959; Rood's Chemistry of Carbon Compounds (edited by S. Coffrey): Elsevier Publishing Company, 1971; and especially Dictionary of Steroids (edited by R.A. Hill, D.N. Kirk, H.L.J. Makin and G.M. Murphy), Chapman & Hall, and this literature is incorporated in the present documentation as a reference. The last publication contains an extensive list of citations of the original works covering the period up to 1990. The present invention also relates specifically to a method for the preparation of (20S) -20 - [(thiomorpholin-4-yl) methyl] 4,4-dimethyl-5a-pregna-8,14-dien-3ß-ol (compound IA), wherein the method comprises the following processing steps: a) starting with (20S) -20-hydroxymethyl -pregna-4-in-3-one; b) introducing two alkyl groups in C4 by alkylation; c) reducing the keto group to a hydroxyl group; d) protecting the resulting hydroxyl group with an acyl group, preferably with a benzyl group; e) introducing a D7 double bond by bromination / dehydrobromination; f) shaping diene D5.7 in diene D8.14 by heating in the presence of acid; g) oxidizing the 20-hydroxyl group to an aldehyde group; h) carrying out the reductive amination of the aldehyde group with thiomorpholine and eliminating the benzyl group by means of a reduction reaction.

The corresponding synthesis scheme of this first synthesis method is presented in Figure 1 (scheme 1). Accordingly, the hydroxyl group on the side chain of (20S) -20-hydroxymethyl-pregna-4-en-3-one 1 is first protected as a silyl ether, for example, as a triisopropylsilyl ether ( TIPS), which results in compound 2 (method step: a). In order to produce compound 3, two methyl groups are introduced by an alkylation with methyl iodide in the presence of a base, such as potassium terbutoxide, at the C4 of the steroid backbone (method step: b). In the next step, the 3-keto group is reduced with a conventional reducing agent, such as lithium aluminum hydride or sodium borohydride (method step: c). The resulting alcohol 4 is subsequently protected, for example, as a benzoate (compound 5, step of the method: d). A second double bond is then introduced through a bromination-dehydrobromination sequence (method step: e). The D5 system of dienes, Resulting in compound 6 is isomerized a posteriori to obtain the diene system D8,14, by heating in the presence of hydrochloric acid, to obtain compound 7 (method step: f). In this acid catalyzed step, only the hydroxyl group in the side chain is deprotected, and compound 7 is obtained. As a result, a selective oxidation of the hydroxyl group in the side chain with Dess-Martin periodinan results in aldehyde 8 ( step of the method: g), which serves as an intermediate to introduce a portion of thiomorpholine in the side chain by reductive amination. For this purpose, various reducing agents can be used, such as sodium borohydride or tris- (acetoxy) borohydride. After the reductive amination, the deprotection of the 3-benzoate is carried out under reducing conditions with ÜAIH4 in a single-vessel process (method step: h). As a result, steroid compound I is obtained according to the present invention. Instead of forming the benzoate in step d) of the method, another acyl protecting group can be introduced, such as, for example, an acetyl group. If acetate is formed instead as an intermediate, the deprotection of the benzoate hydroxyl in step f) of the method occurs not only in the side chain, but also in C3. In this case, a careful selective oxidation of the respective aldehyde must be carried out to avoid oxidation of the hydroxyl group in C3. If benzoate is formed instead of this in step d) of the method, oxidation can only be performed on the hydroxyl group of the side chain. Due to the fact that after step d) of the method the obtained product is crystallized, a simpler purification of the intermediate 8 is possible if a benzoate protecting group is used. In addition, this allows purification in fewer steps, therefore, the formation of the benzoate is preferred. As regards the synthesis of steroid compounds without methyl groups in C4 and / or with another double bond pattern in the steroid backbone, WO 02/079220 A2 can be found, the respective description of which is incorporated by reference.

The examples are provided for the purpose of describing the present invention in greater detail. Example 1: (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-5a-pregna-8,14-dien-3ß-ol (compound IA) a) (20S) -20 -r ((triisopropylsilyl) methyn-preqna-4-en-3-one (Method step: a) To a solution of 30 g of (20S) -20 - [(hydroxymethyl) -pregna- 4-en-3-one and 13.5 g of imidazole in 300 ml of dichloromethane were added 26 ml of triisopropylsilyl chloride by dripping at room temperature.The reaction mixture was stirred for 20 hours at the same temperature and then poured in. The aqueous layer was extracted with ethyl acetate.The organic layers were combined, washed with saline, dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain 45.4 g of (20S) -20 - [((tri-isopropylsilyl) oxy) -methyl] -pregna-4-en-3-one crude as a brown oil, which was used without further purifications MS (CI +): 487 (M + H) b ^ (20S '> -4.4-dimethyl-20-f ((triisoproDysilylipoxy ^ metin-Dreana-5-en-3-one (Method step: b) A solution of 45.4 g of crude (20S) -20 - [((triisopropylsilyl) oxy] methyl] -pregna-4-en-3-one in 320 ml of tetrahydrofuran to a solution of 42.3 g of tert. potassium butylate in 950 ml of tert-butanol at a temperature of 50 ° C. The mixture was stirred for 10 minutes at the same temperature. Then 50 ml of methyl iodide was added and stirring was continued for 1 hour. The reaction mixture was poured into water and extracted with ethyl acetate. The organic layers were combined, washed with saline, dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 27.3 g of (20S) -4,4-dimethyl-20 - [((triisopropylsilyl) oxy) -methyl] -pregna-5-en-3-one as a pale yellow solid. MS (CI +): 515 (M + H) c) (20S) -4,4-dimethyl-20-r ((triisopropylslipoxy) methylene-Dreana-5-en-3ß-ol (Method step: c) To a solution of 27.3 g of (20S) -4,4-d.methyl-20 - [((triisopropylsilyl) oxy) methyl] -pregna-5-en-3-one in 500 ml of tetrahydrofuran is They carefully added 1.24 g of lithium aluminum hydride, in small portions at room temperature, the reaction mixture was stirred for one hour, and then cooled to 0 ° C. Subsequently, 2.5 ml of water was added. water, 2.5 ml of a 1 N solution of sodium hydroxide and 7.5 ml of water The mixture was filtered over celite The filtrate was concentrated under reduced pressure The residue was purified by column chromatography to obtain 18, 2 g of (20S) -4,4-dimethyl-20 - [((triisopropylsilyl) oxy) methyl] -pregna-5-en-3β-ol as a pale yellow solid MS (CI +): 517 (M + H ) d) (20S) -4,4-dimethyl-20-r ((trisopropylsilyl) oxy) methan-preqna-5-en-3ß-ol acetate (Method step: d) To a solution of 18.2 g of (20S) -4; 4-dimethyl- 20 - [((triisopropylsilyl) oxy) methyl] -pregna-5-en-3ß-ol in 175 ml of pyridine was added 6.24 ml of acetic anhydride at room temperature. The reaction mixture was stirred for 20 hours and then poured into an ice / hydrochloric acid mixture. This was extracted with ethyl acetate. The organic layers were combined, washed with saline, dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain 16.2 g of (20S) -4,4-dimethyl-20 - [(( (triisopropylsilyl) oxy) methyl] -pregna-5-en-3-one as a white solid, which was used without further purifications. MS (CI +): 559 (M + H) e) (20S) -4,4-Dimethyl-20- ((triisopropylsilyl) oxy) methyl] -preqna-5,7-diene-3ß-ol acetate (Method step: e) To a solution of 16.2 g of (20S) -4,4-dimethyl-20 - [((triisopropylsilyl) oxy) methyl] -pregna-5-en-3ß-ol acetate in a mixture of 100 ml of benzene and 100 ml of hexane were added 4.93 g of 1,3-dibromo-5,5-dimethylhydantoin in portions at 70 ° C. After 30 minutes, the mixture was cooled to 0 ° C and filtered. The filtrate was evaporated in vacuo. To the resulting residue were added 160 ml of toluene and 7.8 ml of 2,4,6-trimethylpyridine. The mixture was refluxed for 2 hours.,5 hours. After cooling, the reaction mixture was washed with 1N hydrochloric acid, with a saturated solution of sodium bicarbonate and saline. The organic layer was dried over sodium sulfate, filtered and evaporated in vacuo. The residue was purified by column chromatography to obtain 12.5 g of (20S) -4,4-dimethyl-20 - [((tri-isopropylsilyl) oxy) methyl] -pregna-5,7-diene-3ß acetate. -ol as a white solid. MS (CI +): 557 (M + H) ft (20S) -4.4.20-trimethyl-preqna-8.14-dien-3ß.21-diol (Method step: fl A mixture of 16.1 g of ( 20S) -4,4-dimethyl-20- [((trisopropylsilyl) oxl) methyl] -pregna-5,7-diene-3ß-ol, 210 ml of ethanol, 28 ml of benzene and 28 ml of Concentrated hydrochloric acid was refluxed for 6 hours.

After cooling, the mixture was poured into a saturated solution of sodium bicarbonate, extracted with ethyl acetate and washed with saline. The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was recrystallized from dichloromethane and methanol to obtain 4.48 g of (20S) -21-hydroxy-4,4,20-trimethyl-pregna-8,14-diene-3ß-ol. MS (EI +): 358 (M) a) (20S) -3β-hydroxy-4,4,20-trimethyl-preqna-8,14-d-ene-21-al (Method step: q) To a solution of 1 g of (20S) -4,4,20-trimethyl-pregna-8,14-diene-3ß, 21-dol in ml of dichloromethane was added with 5.4 ml of a 0.5 M solution of Dess-Martin at room temperature. The mixture was stirred for one hour, poured into a saturated solution of sodium bicarbonate, extracted with ethyl acetate and washed with saline. The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 230 mg of (20S) -3β-hydroxy-4,4,20-trimethyl-pregna-8,14-diene-21 -al as a white solid. MS (EI +): 356 (M) h) (20V20-f (thiomorpholin-4-methane-4,4-dimethyl-5a-preana-8.14-dien-3ß-ol (Method step: h) 38 were added. mg of sodium tris (acetoxy) borohydride to a solution of 42 mg of (20S) -3β-hydroxy-4,4,20-trimethyl-pregna-8,14-diene-21-al and 20 μl of thiomorpholine in 3 ml of tetrahydrofuran at room temperature The mixture was stirred for two hours, poured into a saturated solution of sodium bicarbonate, extracted with ethyl acetate and washed with saline, the organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure The residue was purified by column chromatography to obtain 15 mg of (20S) -20 - [(thio-morpholin-4-yl) methyl] -4,4-dimethyl-5a-pregna 8,14-dien-3ß-ol as a white solid MS (EI +): 443 (M) The NMR spectra matched the expected structure.The structure was also confirmed with an X-ray structural analysis. Example 2: Reaction according to reaction scheme 1, Fig. 1 It The reactions of Example 1 were repeated to produce the benzoate analogue of intermediate compounds 5, 6, 7 and 8. To this end, the reaction conditions were modified in the various steps of the method as indicated below: Method step: a): TIPSCI, imidazole, CH2Cl2, room temperature, with a reaction time of 4 hours Method step: b): potassium tert-butylate, methyl iodide, tert-butanol, room temperature, with a reaction time of 30 minutes Method step: c): LÍAIH4, tetrahydrofuran, room temperature, with a reaction time of 30 minutes Method step: d): benzyl chloride, pyridine, 0 ° C, with a reaction time from 1 hour; crystallization; 52% in 4 steps Method step: e): 1, 3-dibromo-5,5-dimethyl-imidazolidin-2,4-dione, benzene, hexane, 70 ° C, with a reaction time of 30 minutes; then 2,4,6-trimethylpyridine, toluene, reflux, with a reaction time of 2 hours Method step: f): HCl, ethanol, reflux, with a reaction time of 4 hours, chromatography; 70% in 2 steps Method step: g): Dess-Martin periodinane, CH2CI2, room temperature Method step: h): Tiomorpholine, NaBH (OAc) 3, tetrahydrofuran, room temperature, with a reaction time of 6 hours; then a reaction with LÍAIH4, at room temperature; with a reaction time of 18 hours; chromatography and crystallization from ethanol (2x), yield: 19% in 2 steps, and purification (purity > 93%). The total yield in this reaction sequence was calculated as 6.9% in eight steps.

The following is the reaction sequence and the detailed reaction conditions: ad) benzoate of (20S) -4,4-dimethyl-20-r ((triisopropyl-oxy) metip-preqna-5-en-3ß- ol (compound 5, see the scheme in Fig. 1) (steps of the method: a, b, cvd) This compound was synthesized analogously to the acetate route described in Example 1, the steps to, b, c and d of the method, and is also described in Organic Letters, 5, 1837-1839 (2003). Only crystallization was carried out as a purification step after benzylation. The total yield of steps a, b, c and d was 52%. e) benzoate of (20S) -4,4-dimethyl-20-y ((triisopropylsilyloxy) metill-preqna-5,7-dien-3ß-ol (compound 6. see the scheme in Fig. 1) (Method step: e) To a solution of 30.0 g of (20S) -4,4-dimethyl-20 - [((tri-isopropylsilyl) oxy) methyl] -pregna-5-en-3ß-ol benzoate in a mixture of 160 ml of benzene and 160 ml of hexane were added 9.81 g of 1,3-dibromo-5,5-dimethylhydantoin in portions at 70 ° C. After 30 minutes, the mixture was cooled to 0 ° C. The filtrate was evaporated in vacuo, to the resulting residue were added 280 ml of toluene and 12.7 ml of 2,4,6-trimethylpyridine The mixture was refluxed for 2.5 hours. the reaction mixture was washed with 0.5N hydrochloric acid, with a saturated solution of sodium bicarbonate and saline, the organic layer was dried over sodium sulfate, filtered and evaporated in vacuo to obtain 31.8 g of benzoate. of (20S) -4,4-dimethyl-20 - [((triisopropyl) oxy) methyl] -pregna-5,7-diene-3ß-ol in gross, which was used without further purification. fl (20SV3ß-benzyloxy-4.4.20-trimethyl-preqna-8.14-dien-21-ol (compound 7, see scheme in Fi. 1) (Method step: fl) A mixture of 31.8 was refluxed. g of benzoate of (20S) -4,4-dimethyl-20 - [((triisopropyl) oxl) methyl] -pregna-5,7-diene-3ß-ol, 467 ml of ethanol, 67 ml of benzene and 67 ml of concentrated hydrochloric acid for 5 hours After cooling, the mixture was poured into a saturated solution of sodium bicarbonate, extracted with dichloromethane and washed with saline, the organic layer was dried over sodium sulfate, filtered and The mixture was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 15.7 g of (20S) -3β-benzyloxy-4,4,20-trimethyl-pregna-8,14-diene-21-ol. (20S) -3β-benzyloxy-4.4.20-trimethyl-preqna-8.14-diene-21-al (compound 8, see scheme in Fig. 1) (Method step: q) To a solution of 17 , 6 g of (20S) -3β-benzyloxy-4,4,20-trimethyl-pregna-8,14-dien-21-ol in 470 ml of tetrahydrofuran and 4.1 m pyridine was added with 17.3 ml of Dess-Martin periodinane at 0 ° C. The mixture was stirred for 30 minutes, warmed to room temperature, poured into citrate buffer at pH 7, extracted with dichloromethane and washed with saline. The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to obtain 17.6 g of (20S) -3β-benzyloxy-4,4,20-trimethyl-pregna-8,14-diene-21 -the raw, which was used without additional purifications. h) (20S 20-r (thiomorpholin-4-inmetill-4,4-dimethyl-5-preqna-8,14-dien-3ß-ol (compound I. see the scheme in Fig. 1) (Method step: h) they added 3.9 g of sodium tris (acetoxy) borohydride to a solution of 8.8 g of (20S) -3β-benzyloxy-4,4,20-trimetyl-pregna-8,14-dien-21 - crude and 2.7 ml of thiomorpholine in 400 ml tetrahydrofuran at room temperature The mixture was stirred for two hours, then 10.56 g of lithium aluminum hydride were added in portions, it was set with NaOH and water, it was extracted with ethyl acetate and washed with saline The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure The residue was purified by column chromatography, and two consecutive crystallizations were made from Ethanol to obtain 1.67 g of (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4, -dimethyl-5a-pregna-8,14-dien-3ß-ol as a white solid ( 1, 43 g + 0.24 g, two crystallisation harvests were obtained). these were characterized by 1 H-NMR and MS. The final test of the structure of (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-5a-pregna-8,14-dien-3ß-ol (compound IA) was obtained by performing a structural analysis of X-rays. Chemical characterization v physicochemical, v formulation (solubility): It could be shown that the introduction of a nitrogen-containing side chain improves the solubility of compounds in water (measured by turbometry) . It was found that the solubility of FF-MAS in water was only < 0.1 mg / l.

The solubility of (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-5a-pregna-8,14-dien-3ß-ol (compound IA) in water was determined to be approximately 4 mg / l. Maturation and fertilization of oocytes in vitro (biological evaluation systems) a) Oocyte test enclosed in clusters: Detailed description of the test: It is possible to measure the activation of oocytes arrested in Ml through the disappearance of the germinal vesicle (GV ). The disappearance of the GV, called germinal vesicle degradation (GVB), is followed by the extrusion of the first polar body (PB) (C. Grondahl, JL Ottesen, M. Lessl, P. Faarup, A. Murray, FC Gronvald , C. Hegele-Hartung, I. Ahnfelt-Ronne, "Meiosis-activating sterol promotes resumption of meiosis in mouse oocytes cultured in vitro in contrast to related oxisterols", Biol. Reprod., 58, 1297-1302 (1998); C. Hegele-Hartung, J. Kuhnke, M. Lessl, C. Grondahl, J. Ottesen, HM Beier, S. Eisner, U. Eichenlaub-Ritter, "Nuclear and cytoplasmic maturation of mouse oocytes after treatment with synthetic meiosis-activating sterol in vitro ", Biol. Reprod., 61, 1362-1372 (1999)). The oocysts enclosed in clusters (CEO) were obtained from immature female mice (C57BLJ6J x DBA / 2J F1) weighing 13-16 grams, which were kept under temperature conditions (20-22 ° C), lighting (lights on from 06:00 to 18:00) and relative humidity (50-70%) controlled. The mice received an intra-peritoneal injection of 0.2 ml of gonadotropin containing 20 IU of FSH. 48 hours later, the animals were sacrificed by cervical dislocation. The ovaries were dissected and the oocytes were isolated in Hx medium (see below) under a stereomicroscope, performing the manual rupture of the follicles with a pair of 27-gauge needles. Oocytes enclosed in clusters that presented an intact germ-cell vesicle were placed in an essential minimal medium a (a-MEM without ribonucleosides supplemented with 8 mg / ml of human serum albumin (HSA), 0.23 mM pyruvate, 2 mM glutamine, 100 lU / ml of penicillin and 100 μg / ml streptomycin). To maintain the oocysts in the germinal vesicle stage, this medium is supplemented with 3 mM hypoxanthine, called Hx medium.

Dilutions of (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-5a-pregna-8,14-dien-3ß-ol (compound IA) were prepared from a stock solution containing 1 mg / ml dissolved in ethanol. The addition of ethanol (3.8 μl / ml) to the controls did not affect the control level. The CEOs (35-45 CEO in 0.4 ml of Hx medium) were grown in a humidified atmosphere of 5% CO2 in air for 24 hours at 37 ° C. A control cavity (identical medium without the addition of the test compound) was always cultivated simultaneously with the control cavities. At the end of the culture period, the amount of CEO was counted with germ vesicles (GV), degraded germ vesicles (GVB) and polar bodies (PB), respectively. The% of GVB, defined as the percentage of CEO that experienced GVB, with respect to the total amount of CEO in said cavity, was calculated as: GVB = (amount of GVB + amount of PB / total amount of CEO) x 100% PB was defined as the percentage of CEO who presented an extruded polar body, with respect to the total number of CEOs in said cavity. Results: In the Flg. 2 the activation of meiosis with (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-5a-pregna-8,14-dien-3ß-ol (compound IA) in CEO, monitored through the GVB and PB (n = 5; * p < 0.05). The new compound IA significantly improved the activation of meiosis at concentrations of 1 μM (p < 0.01; n = 10), 3 μM (p <0.05, n = 7) and 10 μM (p <0.001, n = 6). The same experiments were repeated with the other new compounds: IB: (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-5a-pregna-8 (14) -en-3ß-ol IC: (20S) -20 - [(thiomorpholin-4-yl) methyl] -5a-pregna-8,14-dien-3ß-ol ID: (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-pregna-5,7-diene-3ß-ol and IE: (20S) -20 - [(thiomorpholin-4-yl) methyl] -5a-pregna-8 (14) -en-3ß -ol. The results of the activation of meiosis in the presence of hypoxanthine (Hx) in CEO are given in Table 1. Two experiments were performed for each compound, and the results were purchased with those obtained with Hx medium as control and FF-MAS at a concentration of 10 μM. In addition to concentration dependence, the activation of meiosis in the presence of Hx in CEO was determined using compound IE, which, in addition to compound IA, was the most efficient compound evaluated. Table 2 shows the dependence of the concentration of the activation of meiosis, in comparison with that obtained with the medium Hx, FF-MAS and (20S) -20 - [(piperidin-1-yl) methyl] -4 , 4-dimethyl-5a-pregna-8,14-dien-3ß-ol (described in WO 02/079220 A2 as the compound No. 2). b) In vitro fertilization (IVF) To investigate the effect of (20S) -20 - [(thomorpholin-4-yl) methyl] -4,4-dimethyl-5a-pregna-8,14-dien- 3ß-ol (compound IA) on the IVF rate, CEO in vitro was matured in the presence or absence of this new compound, and then the IVF was performed. FF-MAS and (20S) -20 - [(piperidin-1-yl) methyl] -4,4-dimethyl-5a-pregna-8,14-diene-3ß-ol (described in WO 02/079220 A2) were used. as compound No. 2) as control compounds. As the means for in vitro maturation in a clinical preparation would not contain hypoxanthine, this new compound was not used in any medium, which practically resulted in 100% activation of meiosis in the oocytes, due to spontaneous meiotic maturation. Accordingly, subsequent changes in IVF are likely due to the effects of the test compounds on cytoplasmic maturation. Immature female mice (C57B1J6J x DBA / 2J F1) with an age of 21-24 days were used as oocyte donors. The animals were administered an i.p. of 10 IU pregnant mare serum gonadotropin (PMSG) to induce follicular growth. 48 hours later, the animals were sacrificed by cervical dislocation and the ovaries were dissected. As sperm donors, eight-week-old female mice (C57BL / 6J x DBA / 2J F1) were used.

Oocytes were isolated from the ovaries by manually breaking the leaflets with a pair of 27-gauge needles. Naked spherical oocysts (NkO) and CEO were selected that had an intact GV, and were placed in a minimum essential medium a ( α-MEM without ribonucleosides, Gibco BRL, Gaithersburg; Cat # 22561) supplemented with 8 mg / ml human serum albumin (HSA), 0.23 mM pyruvate, 2 mM glutamine, 100 μU / ml penicillin and 100 μg / ml streptomycin. The oocytes were washed three times in culture medium without oil coating, in 4 cavity multiple cavity plates containing 0.4 ml of the respective oocyte culture medium. The oocytes were cultured at 37 ° C in a humidified atmosphere of 5% CO 2, in air in the presence of (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dιmethyl- 5a-pregna-8,14-d-ene-3ß-ol (compound IA), FF-MAS and (20S) -20 - [(piperidin-1-yl) methyl] -4,4-dimethyl-5a -pregna-8,14-dien-3ß-ol, where the latter constituted comparison compounds. The oocyte culture medium is supplemented with 6.4% fetal bovine serum (FBS). Solutions of the new compound IA and the two mentioned comparison compounds were prepared from a stock solution containing 1 mg / ml of compound dissolved in absolute ethanol. The control group was cultured with a corresponding amount of ethanol. After a culture period of 18-19 hours, the oocytes were further processed for in vitro fertilization. Only the oocytes that presented GVB or PB were defined as "mature oocytes in vitro". The oocytes were briefly washed without any test compound, and then they were transferred to insemination plates prepared in advance, containing approximately 600,000 trained sperm cells per 500 μl of IVF medium (obtained from the epididymis of male mice). The plates were then incubated under defined gas conditions (5% CO2 in air) at 37 ° C, in a modified IVF a-MEM medium, supplemented with 8 mg / ml HSA, 0.23 mM pyruvate, 100 IU of penicillin / ml and 100 μg of streptomycin / ml, as previously described for oocyte maturation. An oocyte examination was performed 22-24 hours after insemination to verify fertilization and record the number of pronuclei (PN) and embryos in the 2-cell stage. The fertilization rate was determined as the number of oocytes that reached the PN or embryo stage of 2 cells, compared to the total amount inseminated. Results: Fig. 3 shows higher fertilization rates of mouse oocytes with the new compound IA and the two mentioned compounds, (20S) -20 - [(piperidin-1-yl) methyl] -4,4 -dimethyl-5a-pregna-8,14-dien-3ß-ol (compound No. 2) or FF-MAS, compared to control vehicle (* p <; 0.05; *** p < 0.001; n = 18). Fig. 3a shows the values, expressed as a percentage of the total oocytes used in the experiment (approximately 100 oocytes per group per experiment). Fig. 3b shows the data for the stimulation factor, normalized according to the vehicle group. Re-transfer study: In vitro fertilization was performed as previously described for the IVF experiment. One day before the re-transfer, the female recipient mice (adoptive mothers) were crossed in a proestrus cycle with vasectomized males, in order to trigger pseudopregnancy. A day later, a vaginal verification was performed. Animals with a positive, sperm-free vaginal plug (successful crossover) entered the study. This day was called day 0 of gestation (= day 0 after sterile copulation, day 0 p.c.). Eighteen embryos from 2 cells were transferred to the right tube of the adoptive mothers on day 0 after sterile copulation, under anesthesia. The contralateral tube was not used (pseudopregnancy verification). The adoptive mothers sacrificed on the 19th day of gestation. The fetuses were extracted and examined. The ovaries and uteri were removed to record the number of implantation sites, the viable and dead fetuses, and the reabsorption. The apparently non-pregnant uteri were placed in an aqueous solution of 10% ammonium sulphide for approximately 10 minutes to color possible sites of implantation in the endometrium. The viable fetuses were weighed individually, and their sex was determined by inspecting their gonads.

In the fertility test, 269 mice were distributed in 6 groups at random, and they were implanted with 7-10 eggs fertilized in vitro. In vitro fertilization took place in the presence of a vehicle (group 1) or a substance (groups 2 to 6). The groups were distributed as indicated in Table 3. The effect of increasing the fertility of the substances, as compared to the control vehicle, was of great interest. Moreover, an improvement was also investigated, compared to the substance FF-MAS. The variables of primary interest are the number of implantation sites and the number of viable fetuses. Methods: It is necessary to consider that implantation data, as well as viable fetuses, must have a binary character, that is, they must be able to express themselves in rates or in a number of successes and failures with respect to the quantity of eggs transferred. Accordingly, the methods detailed by D. Collett: "Modeling Binary Data", 2nd edition, Chapman & Hall / CRC, Boca Ratón, constitute appropriate approaches for data analysis. As the probabilities of success in a group, compared to the probabilities of success in another group, are appropriate for the interpretation of these data, the analysis focuses on the computation of probability relationships. The likelihood ratio of one group, compared to the other, is defined as: Odds ratio (group 1, group 2) = or?, 2 = probability (group 1) / probability (group 2), where the probabilities of a group can be estimated from the number of successes, divided by the number of failures in that group. To obtain significant results in all cases, the 95% confidence intervals are computed for probabilities such as: [exp (log (or?, 2) /1.96 - se (log (or?, 2))); exp (log (on, 2) + 1.96 - se (log (or?, 2)))], where se (log (or1? 2)) is the standard error of the probability relation (see D.

Collett (2003) to find the corresponding formula). It can be established that the probabilities of one group are significantly greater than the probabilities in the other group if the lower confidence limit is greater than 1. Comparisons with the control vehicle: As a basis for computing the probability relationships, the number of successes is computed (ie, the number of implantation sites for the number of viable fetuses) and failures (ie, the number of eggs transferred minus the number of implantation sites or the number of viable fetuses) for all groups. Table 4 shows these values, and the resulting probabilities for the six groups. Table 5 shows the probability relationships of the five groups, as compared to the control vehicle, and their confidence intervals.

For the number of implantation sites, compound IA shows a significant improvement in the probabilities in both doses. This new compound, at a concentration of 1 μM, increases the chances of successful implantation by 51%, while a concentration of 10 μM leads to a 55% better chance of successful implantation. Neither FF-MAS nor compound No. 2 ((20S) -20 - [(piperidin-1-yl) methyl] -4,4-d, methyl-5a-pregna-8,14-d-ene-3a- ol) significantly improve the odds. Compound IA increases the chances of success, in terms of viable fetuses, by 70%, using the lowest doses. None of the other groups present significant differences with respect to the control vehicle. Comparisons with FF-MAS: Table 6 shows the probability relationships of groups 3 to 6, compared to FF-MAS (group 2), and their confidence intervals. Since all confidence intervals include the value 1, it can not be concluded that there are significant differences between the four groups of substances and FF-MAS. Fig. 4 shows the implantation rate for the embryos of 2 transferred cells, which reaches higher values for the treatment with FF-MAS (10 μM) and the new compounds IA, at concentrations of 1 μM and 10 μM. The number of live embryos is represented in Fig. 5. Again, the numbers of live embryos were recorded after treatment with FF-MAS and the new compound IA (1 μM and 10 μM). The pregnancy rates for each group are presented in Fig. 6. Notably, the new compound IA has a beneficial effect on the embryo rate of 2 cells and blastocysts at a concentration of 0.1, an effect that tends to be even greater at a concentration of 1 μM. It will be understood that the examples and embodiments described in this documentation are provided for illustrative purposes only, and that those skilled in the art may suggest various modifications and changes taking them as a reference, as well as combinations of features described in this application, all of which they will be included in the spirit and scope of the described invention and the appended claims. All publications, patents and patent applications cited in this documentation are incorporated by reference.

Table 1: Activation of meiosis in the presence of Hx in CEO *) (20S) -20 - [(piperidin-1-yl) methyl] -4,4-dimethyl-5a-pregna-8, 14-dien-3ß-ol Table 3: *) (20S) -20 - [(piperidin-1-yl) methyl] -4,4-dimethyl-5a-pregna-8,14-dien-3ß-ol Table 4: Successes, failures and probabilities per group *) (20S) -20 - [(piperidin-1-yl) methyl] -4,4-d-methyl-5a-pregna-8,14-dien-3ß-ol Table 5: Probability relationships and confidence limits for comparison with the control vehicle *) (20S) -20 - [(piperidin-1-yl) metl] -4,4-d-methyl-5a-pregna-8,14-di-3-yl Table 6: Relationships of probabilities and confidence limits for comparison with FF-MAS t) (20S) -20 - [(piperidin-1-yl) methyl] -4,4-dimethyl-5a-pregna-8,14-dien-3ß-ol

Claims (12)

1. CLAIMS 1. A steroid thiomorpholino compound of general formula I where, in the I 'portion of compound I r each link between C5 and C6, between C6 and C7, between C7 and C8, between C8 and C9, between C8 and C14, and between C14 and C15, independently, is a single link or a double link, where at least one of these links is a double bond, taking into account that there are no double bonds in the steroid skeleton, exclusively between C5 and C6, and where R4 and R4, independently, are selected from the group comprising hydrogen and methyl.
2. 2. The steroid compound according to claim 1, wherein in the portion with the general formula I 'there is a double bond present between C8 and C14, or there are two double bonds present between C8 and C9, and between C14 and C15, or there are two double bonds present between C5 and C6, and between C7 and C8.
3. 3. The steroid compound according to any of claims 1 and 2, which is selected from the group comprising: (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-5a -pregna-8,14-dien-3ß-ol: IA (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-5a-pregna-8 (14) -en-3ß-ol: IB (20S) -20 - [(thiomorpholin-4-yl) methyl] -5a-pregna-8,14-dien-3ß-ol: IC (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-pregna-5,7-diene-3ß-ol ID (20S) -20 - [(thiomorpholin-4-yl) methyl] -5a-pregna-8 (14) -en-3ß-ol: IE
4. 4. A pharmaceutical composition comprising at least one steroid thiomorpholino compound of general formula I according to any of claims 1-3 and at least one excipient acceptable for pharmaceutical use.
5. 5. The pharmaceutical composition according to claim 4, wherein the steroid compound of general formula I is included in an effective amount.
6. 6. A use of the steroid thiomorpholino compound of general formula I according to any of claims 1-3, which is for the preparation of a pharmaceutical composition useful for regulating reproduction, especially meiosis. The use according to claim 6, which is for an application that is not in vivo. 8. A use of the steroid thiomorpholino compound of general formula I according to any of claims 1-3, which is for the preparation of a contraceptive or a pro-fertility drug. 9. A method for regulating reproduction, especially meiosis, comprising administering to a subject in need of such regulation an effective amount of at least one steroid thiomorpholino compound of general formula I according to any of claims 1-3. 10. A method for improving the possibility of an oocyte developing in a mammal, comprising contacting an oocyte extracted from the mammal with the steroid thiomorpholino compound according to any of claims 1-3. 11. A method for the preparation of (20S) -20 - [(thiomorpholin-4-yl) methyl] -4,4-dimethyl-5a-pregna-8,14-dien-3ß-ol, which comprises a) starting with (20S) -20-hydroxymethyl-pregna-4-en-3-one; b) introducing two alkyl groups in C4 by alkylation; c) reducing the keto group to a hydroxyl group; d) protecting the resulting hydroxyl group with an acyl group, preferably with a benzyl group; e) introducing a D7 double bond by bromination / dehydrobromination; f) isomerizing the D5.7 diene in the D8,14 diene by heating in the presence of acid; g) oxidizing the 17-hydroxyl group in an aldehyde group; h) carrying out the reductive amination of the aldehyde group with thiomorpholine and eliminating the benzyl group by means of a reduction reaction. The method according to claim 11, wherein the acyl group is a benzoate group.